Bakhetia Manjula, Charlton Wayne, Atkinson Howard J, McPherson Michael J
Centre for Plant Sciences, Faculty of Biological Sciences, University of Leeds, UK.
Mol Plant Microbe Interact. 2005 Oct;18(10):1099-106. doi: 10.1094/MPMI-18-1099.
RNA interference (RNAi) is a powerful tool for the analysis of gene function in model organisms such as the nematode Caenorhabditis elegans. Recent demonstrations of RNAi in plant parasitic nematodes provide a stimulus to explore the potential of using RNAi to investigate disruption of gene function in Meloidogyne incognita, one of the most important nematode pests of global agriculture. We have used RNAi to examine the importance of dual oxidases (peroxidase and NADPH oxidase), a class of enzyme associated with extracellular matrix cross-linking in C. elegans. RNAi uptake by M. incognita juveniles is highly efficient. In planta infection data show that a single 4-h preinfection treatment with double-stranded RNA derived from the peroxidase region of a dual oxidase gene has effects on gene expression that are phenotypically observable 35 days postinfection. This RNAi effect results in a reduction in egg numbers at 35 days of up to 70%. The in vitro feeding strategy provides a powerful tool for identifying functionally important genes, including those that are potential targets for the development of new agrochemicals or transgenic resistance strategies.
RNA干扰(RNAi)是分析线虫秀丽隐杆线虫等模式生物中基因功能的强大工具。最近在植物寄生线虫中对RNAi的证明促使人们探索利用RNAi研究全球农业中最重要的线虫害虫之一南方根结线虫基因功能破坏的潜力。我们利用RNAi研究了双氧化酶(过氧化物酶和NADPH氧化酶)的重要性,这是一类与秀丽隐杆线虫细胞外基质交联相关的酶。南方根结线虫幼虫对RNAi的摄取效率很高。植物体内感染数据表明,用来自双氧化酶基因过氧化物酶区域的双链RNA进行单次4小时感染前处理,对基因表达有影响,在感染后35天可观察到表型变化。这种RNAi效应导致在35天时卵数量减少高达70%。体外喂食策略为鉴定功能上重要的基因提供了强大工具,包括那些作为新型农用化学品或转基因抗性策略开发潜在靶点的基因。